Spectroscopic imaging combines digital imaging and molecular spectroscopy techniques, which can include Raman scattering, fluorescence, photoluminescence, ultraviolet, visible and infrared absorption spectroscopies. When applied to the chemical analysis of materials, spectroscopic imaging is commonly referred to as chemical imaging. Instruments for performing spectroscopic (i.e. chemical) imaging typically comprise an illumination source, image gathering optics, focal plane array imaging detectors and imaging spectrometers.
In general, the sample size determines the choice of image gathering optic. For example, a microscope is typically employed for the analysis of sub micron to millimeter spatial dimension samples. For larger objects, in the range of millimeter to meter dimensions, macro lens optics are appropriate. For samples located within relatively inaccessible environments, flexible fiberscope or rigid borescopes can be employed. For very large scale objects, such as planetary objects, telescopes are appropriate image gathering optics.
For detection of images formed by the various optical systems, two-dimensional, imaging focal plane array (FPA) detectors are typically employed. The choice of FPA detector is governed by the spectroscopic technique employed to characterize the sample of interest. For example, silicon (Si) charge-coupled device (CCD) detectors or CMOS detectors are typically employed with visible wavelength fluorescence and Raman spectroscopic imaging systems, while indium gallium arsenide (InGaAs) FPA detectors are typically employed with near-infrared spectroscopic imaging systems.
Spectroscopic imaging of a sample can be implemented by one of two methods. First, a point-source illumination can be provided on the sample to measure the spectra at each point of the illuminated area. Second, spectra can be collected over the entire area encompassing the sample simultaneously using an electronically tunable optical imaging filter such as an acousto-optic tunable filter (“AOTF”) or a LCTF. This may be referred to as “wide-field imaging”. Here, the organic material in such optical filters are actively aligned by applied voltages to produce the desired bandpass and transmission function. The spectra obtained for each pixel of such an image thereby forms a complex data set referred to as a hyperspectral image (“HSI”) which contains the intensity values at numerous wavelengths or the wavelength dependence of each pixel element in this image.
Spectroscopic devices operate over a range of wavelengths due to the operation ranges of the detectors or tunable filters possible. This enables analysis in the Ultraviolet (“UV”), visible (“VIS”), near infrared (“NIR”), short-wave infrared (“SWIR”), mid infrared (“MIR”) wavelengths and to some overlapping ranges. These correspond to wavelengths of about 180-380 nm (UV), 380-700 nm (VIS), 700-2500 nm (NIR), 9850-1800 nm (SWIR), and 650-1100 nm (MIR), 1200-2450 nm (LWIR).
When operating in these wavelength ranges, certain objects will become visible in the resulting spectroscopic image. These objects may be referred to as “active” objects. Other “inactive” objects may not be visible in the spectroscopic image. It would be advantageous if a method would enable quick and reliable assessment of these “inactive” objects. This would enable visualization of previously invisible objects.
In certain situations, it may be advantageous to combine spectroscopic images with RGB or white light images of a region of interest to provide a more robust assessment of objects present in a field of view. It may be particularly advantageous to combine white light images with spectroscopic images when using a microscope based imaging system to analyze samples comprising unknown objects. The current state of the art requires using multiple optical devices or channels to generate these different types of images. Using multiple optical devices is expensive and time consuming. There exists a need for a method that would enable the generation of different types of images without requiring multiple optical devices.